Activity checks performed on D64V IN variations produced in E. coli demonstrated they had no strand ubiquitin conjugating transfer exercise, and their genes are, therefore, safe to make use of in immunization. All three integrase variants were highly expressed in human and murine cells. The amount of eukaryotic expression achieved 700 pg per cell, exceeding the levels observed for that virus taken HIV 1 enzyme genes by almost 50-fold. None of the mutations had any impact on the degree of IN expression. Ergo, the humanized IN genes met all requirements set for the effective gene immunogens. It was confirmed by the outcome of the IN gene immunization of BALB/c rats. All three IN genes were clearly immunogenic for mouse T-cells. CD8 and CD4 T-cell responses were mainly directed against a cluster of epitopes at aa 209 239 of IN. IFN c/IL 2 response of murine PBMC against this cluster was registered currently on day 15 after immunization. By morning 27, T cell responses of splenocytes to stimulation with IN209 and MIN219 had somewhat pyridine extended. IN aa 209 239 of opinion HIV 1 clade An appeared to contain a murine T cell epitope. A solid T cell response against this region induced by all IN gene versions suggested its use as a cause epitope to monitor integrasespecific T cell responses. Recognition of other peptides representing mouse and human T-cell epitopes nearby at aa 66 98 and 169 190 was weak and occurred primarily in the shape of IL 2 production. T-cell stimulation by IN derived peptides was further analyzed by FACS. In all groups obtaining IN genes, stimulation by the pool of proteins representing mouse CD4 and specific Hedgehog inhibitor CD8 T cell epitopes induced production of IFN h, IL 2, and/or TNF a by 0. 08 to 0. Fortnight CD4 cells, of IFNc or TNF a by 0. 8 to 1. 62-room CD8, and of IL 2 by 0. A day later CD8 T cells. None of the stimulated T cells produced IL 4. IFN h is the most often measured cytokine related to protection against viral infections. Thus, all three artificial IN genes behaved as effective gene immunogens able to cause potent Th1 type reactions in both CD8 and CD4 T cells. Release of both TNF and IFNc a by effector CD8 T cells is critically essential for protection against viral infections. IL 2 supports the secondary growth of memory CD8 T cells and generation of the long run protective immunity,. Technology of most three cytokines is considered to be a requisite for an efficient antiviral immunization. Production of cytokines is hierarchical in character: a lot of the epitope certain CTLs make IFN c, some, IFN c TNF a, and still a smaller subset, referred to as polyfunctional, all three cytokines,. Polyfunctional T-cells have already been associated with a highly effective control of intracellular infections, specifically of viral replication, and with strong protection in vaccination,,,,,.